Culture Substrate Research Articles

Transformative Biomaterials in Burn Care: Utilizing Scaffolds, Hydrogels, and Chitosan for Optimized Healing and Scar Reduction

With over a million patients per year in the USA alone, burns are among the most common injuries in the world. In addition to preventing difficulties during burn injuries, the growing impact of scaffolds, hydrogels, cell scaffolds, and cell sheets with healing boosting elements expedite triggers and strengthens re-epithelialization and wound healing, which limits the creation of scars. While superficial partial-thickness and superficial burns typically heal without surgery, severe burns require special care, including topical antimicrobial surgery or bandages. Usually composed of polymeric biomaterials, scaffolds offer the structural support needed for cell adhesion and the subsequent formation of tissue. The term "tissue engineering triad" often refers to cells, scaffolds, and growth-stimulating signals. This may be accomplished in several ways by combining different polymers. Simple (planar) 3D structures may be produced using conventional and rapid prototyping techniques, but complicated structures have been successfully produced by carefully controlling the Molds and processing parameters. For biomedical and tissue engineering applications, chitosan has been widely employed (skin, bone, cartilage, and vascular grafts to substrates for mammalian cell culture). Additionally, it is renewable, biocompatible, biodegradable, nonantigenic, bioactive, and nontoxic.

Thermoresponsive Brush Coatings for Cell Sheet Engineering with Low Protein Adsorption above the Polymers' Phase Transition Temperature.

Thermoresponsive polymer coatings on cell culture substrates enable noninvasive cell detachment and cell sheet fabrication for biomedical applications. Optimized coatings should support controlled culture and detachment of various cell types and allow chemical modifications, e.g., to introduce specific growth factors for enhanced gene expression. Furthermore, the sterilization and storage stability of the coatings must be assessed for translational attempts. Poly(glycidyl ether) (PGE) brush coatings with short alkoxy side chains provide a versatile platform for cell culture and detachment, but their polyether backbones are susceptible to oxidation and degradation. Thus, we rationally designed potential alternatives with thermoresponsive glycerol-based block copolymers comprising a stable polyacrylate or polymethacrylate backbone and an oligomeric benzophenone (BP)-based anchor. The resulting poly(ethoxy hydroxypropyl acrylate-b-benzophenone acrylate) (pEHPA-b-BP) and poly(ethoxy hydroxypropyl methacrylate-b-benzophenone methacrylate) (pEHPMA-b-BP) block copolymers preserve the short alkoxy-terminated side chains of the PGE derived structure on a stable, but hydrophobic, aliphatic backbone. The amphiphilicity balance is maintained through incorporated hydroxyl groups, which simultaneously can be used for chemical modification. The polymers were tailored into brush coatings on polystyrene surfaces via directed adsorption using the BP oligomer anchor. The resulting coatings with thickness values up to ∼3 nm supported efficient adhesion and proliferation of human fibroblasts despite minimal protein adsorption. The conditions for cell sheet fabrication on pEHPA-b-BP were gentler and more reliable than on pEHPMA-b-BP, which required additional cooling. Hence, the stability of pEHPA-b-BP and PGE coatings was evaluated post gamma and formaldehyde (FO) gas sterilization. Gamma sterilization partially degraded PGE coatings and hindered cell detachment on pEHPA-b-BP. In contrast, FO sterilization only slowed detachment on PGE coatings and had no adverse effects on pEHPA-b-BP, maintaining their efficient performance in cell sheet fabrication.

Optimization of Magnetic Biochar Preparation Process, Based on Methylene Blue Adsorption

The search for low-cost and effective adsorbents for the removal of organic dyes from contaminated water is urgently needed. The substantial amount of waste mushroom cultivation substrates generated in practical production can serve as an ideal material for the preparation of adsorbents. In this study, we investigated the main control parameters affecting the performance of magnetic mushroom substrate biochar and optimized the process of preparing biochar by using the Plackett–Burman and central composite design methods. Various analytical techniques including SEM, EDX, BET, and VSM were used to characterize the biochar. The results indicate that the carbonization temperature had the most significant impact on the yield and adsorption performance of biochar. Under the conditions of a carbonization temperature of 600 °C, a carbonization retention time of 1 h, and an impregnation ratio of 0.1, the yield and methylene blue adsorption value of magnetic biochar were 42.54% and 2297.04 μg/g, respectively, with a specific surface area of 37.17 m2/g. This biochar effectively removed methylene blue from the solution, demonstrating a high economic efficiency for wastewater treatment and pollution control. Furthermore, the adsorption–desorption cycle studies revealed its excellent stability and reusability. Additionally, based on the response surface methodology, a three-dimensional surface model of the adsorption performance of magnetic biochar under different carbonization conditions was established, providing a theoretical basis for the preparation of magnetic biochar from agricultural wastes.

Biosensor for integrin inhibition of mammalian cell adhesion and migration using micropatterned cell culture substrate and retroreflective optical signaling

Integrins are a family of transmembrane receptors that play a crucial role in cell adhesion and migration. Integrins can uniquely transduce biochemical signals bidirectionally across the membrane and physically link the cell-cell and cell-extracellular matrix (ECM) with ligand bonds. The arginyl-glycyl-aspartic acid (RGD) peptide motif is present in the ECM as a minimal recognition sequence for integrins. To leverage this property in cell-based therapy, RGD variants, such as cyclic-type RGDfK (c(RGDfK)), which share a similar structure with RGD but exhibit a higher affinity for integrins, have been developed. However, because most evaluation methods for newly developed RGD variants focus on affinity strength, tools for cellular effects are required. In this study, we developed a new platform that integrates micropatterned three-dimensional cell culture substrates with a non-spectroscopic optical analysis system to quantitatively analyze the effects of RGD variants on cell adhesion and migration. The specially micropatterned substrate provides a cell adhesive and migration area to provide a restricted analysis area. Owing to the characteristics of retroreflective Janus particles (RJPs), a non-spectroscopic optical analysis system provides long-term stable optical verification properties and a simple optical setup. These techniques were integrated to quantitatively determine the integrin inhibitory effect of various concentrations of RGD variant. To demonstrate the efficacy of the developed cellular level RGD variant testing platform, the model cell line L929 fibroblast and model RGD variant c(RGDfK) were analyzed ranging from 0 to 10 μM. The results showed that the developed system could effectively and quantitatively analyze the effects of RGD variants on cells across various concentrations.

Using Polycaprolactone Nanofibers for the Proof-of-Concept Construction of the Alveolar-Capillary Interface.

The alveolar-capillary interface is the key functional element of gas exchange in the human lung, and disruptions to this interface can lead to significant medical complications. However, it is currently challenging to adequately model this interface invitro, as it requires not only the co-culture of human alveolar epithelial and endothelial cells but mainly the preparation of a biocompatible scaffold that mimics the basement membrane. This scaffold should support cell seeding from both sides, and maintain optimal cell adhesion, growth, and differentiation conditions. Our study investigates the use of polycaprolactone (PCL) nanofibers as a versatile substrate for such cell cultures, aiming to model the alveolar-capillary interface more accurately. We optimized nanofiber production parameters, utilized polyamide mesh UHELON as a mechanical support for scaffold handling, and created 3D-printed inserts for specialized co-cultures. Our findings confirm that PCL nanofibrous scaffolds are manageable and support the co-culture of diverse cell types, effectively enabling cell attachment, proliferation, and differentiation. Our research establishes a proof-of-concept model for the alveolar-capillary interface, offering significant potential for enhancing cell-based testing and advancing tissue-engineering applications that require specific nanofibrous matrices.

Optimizing cultivation substrate moisture and calcium application for the comprehensive growth of cucumber under different air temperatures

Optimizing cultivation substrate moisture and calcium application for the comprehensive growth of cucumber under different air temperatures

Effects of Clump Size on the Pluripotency and Proliferation in the Passaging Process of Mouse Induced Pluripotent Stem Cells

Induced pluripotent stem cells (iPSCs) are a promising cell source because of their pluripotency and self-renewal abilities. However, there is a risk of pluripotency loss during cell expansion. Particularly, cell passaging is associated with a higher risk of decreasing cell quality. There are two iPSC passaging methods: single-cell and clump passaging. Single-cell passaging is a rapid and simple method for cell manipulation, whereas clump passaging is superior for maintaining iPSC pluripotency. Therefore, clump passaging is a robust method for expanding iPSCs while maintaining their pluripotency. However, clump size control during clump passaging is difficult because colony fragmentation is performed manually by pipetting the colonies detached from the cell culture substrates. In this study, the effect of pipetting on iPSC colony fragmentation was evaluated and the relationship between iPSC clump size and pluripotency was clarified. An automated pipetting device was developed to standardize the clump passage process. The effect of clump size on the pluripotency and proliferative capacity of mouse iPSCs was investigated. Clump size was controlled by varying the number of pipetting cycles, and pluripotency and proliferation were assessed via alkaline phosphatase staining and flow cytometry. Our results revealed that a decrease in clump size corresponded to an increase in cell proliferation, while pluripotency maintenance was optimized under specific clump sizes. These results underscore the significance of clump size for stem cell quality, emphasizing the need for a balanced approach to maintain pluripotency while fostering proliferation in the cell expansion culture for iPSCs.

Bacterial Acyl Homoserine Lactones Triggered Non‐Native Substrate Hydroxylation Catalyzed by Directed‐Evolution‐Derived Cytochrome P450BM3 Mutants

AbstractWe report the directed evolution of cytochrome P450BM3 to efficiently utilize the bacterial quorum sensing signalling molecule N‐decanoyl homoserine lactone (C10‐HSL) as an effective decoy molecule. This represents the first important step in our endeavor to develop of a self‐sufficient decoy‐molecule system in whole‐cells that only necessitates the addition of culture medium and substrate to realize the hydroxylation of non‐native substrates. Following five rounds of directed evolution, mutant P450BM3, in the presence of C10‐HSL, catalyzed the hydroxylation of benzene at a rate of 475 min−1, the highest turnover rate recorded for any P450 enzyme, and achieving a 46% yield in a whole‐cell reaction system. High‐resolution X‐ray crystal structure analysis of a series of mutants narrates the directed evolution process, revealing how C10‐HSL is fixed in the binding pocket to permit binding of non‐native substrates. Finally, introduction of the C10‐HSL synthase gene ExpI into Escherichia coli, enabled the in situ production of C10‐HSL, realizing, for the first time, the hydroxylation of non‐native substrates without the need for the laborious synthesis and addition of decoy molecules.

Rescue of mitochondrial dysfunction through alteration of extracellular matrix composition in barth syndrome cardiac fibroblasts

Fibroblast-ECM (dys)regulation is associated with a plethora of diseases. The ECM acts as a reservoir of inflammatory factors and cytokines that mediate molecular mechanisms within cardiac cell populations. The role of ECM-mitochondria crosstalk in the development and progression of cardiac disorders remains uncertain. We evaluated the influence of ECM produced by stromal cells from patients with the mitochondrial cardiomyopathy (Barth syndrome, BTHS) and unaffected healthy controls on cardiac fibroblast (CF) metabolic function. To do this, cell-derived matrices CDMs were generated from BTHS and healthy human pluripotent stem cell-derived CFs (hPSC-CF) and used as cell culture substrates. BTHS CDMs negatively impacted the mitochondrial function of healthy hPSC-CFs while healthy CDMs improved mitochondrial function in BTHS hPSC-CFs. Mass spectrometry comparisons identified 5 matrisome proteins differentially expressed in BTHS compared to healthy CDM. Our results highlight a key role for the ECM in disease through its impact on mitochondrial function.

Metabolome and Metagenome Integration Unveiled Synthesis Pathways of Novel Antioxidant Peptides in Fermented Lignocellulosic Biomass of Palm Kernel Meal.

Approximately one-third of the entire world's food resources are deemed to be wasted. Palm kernel meal (PKM), a product that is extensively generated by the palm oil industry, exhibits a unique nutrient-rich composition. However, its recycling is seldom prioritized due to numerous factors. To evaluate the impact of enzymatic pretreatment and Lactobacillus plantarum and Lactobacillus reuteri fermentation upon the antioxidant activity of PKM, we implemented integrated metagenomics and metabolomics approaches. The substantially enhanced (p < 0.05) property of free radicals scavenging, as well as total flavonoids and polyphenols, demonstrated that the biotreated PKM exhibited superior antioxidant capacity. Non-targeted metabolomics disclosed that the Lactobacillus fermentation resulted in substantial (p < 0.05) biosynthesis of 25 unique antioxidant biopeptides, along with the increased (p < 0.05) enrichment ratio of the isoflavonoids and secondary metabolites biosynthesis pathways. The 16sRNA sequencing and correlation analysis revealed that Limosilactobacillus reuteri, Pediococcus acidilactici, Lacticaseibacillus paracasei, Pediococcus pentosaceus, Lactiplantibacillus plantarum, Limosilactobacillus fermentum, and polysaccharide lyases had significantly dominated (p < 0.05) proportions in PMEL, and these bacterial species were strongly (p < 0.05) positively interrelated with antioxidants peptides. Fermented PKM improves nutritional value by enhancing beneficial probiotics, enzymes, and antioxidants and minimizing anti-nutritional factors, rendering it an invaluable feed ingredient and gut health promoter for animals, multifunctional food elements, or as an ingredient in sustainable plant-based diets for human utilization, and functioning as a culture substrate in the food sector.

Growth, physiological and N, P, K accumulation responses of Erythropalum scandens Bl. Seedlings under different substrates

Erythropalum scandens Bl. is a medicinal woody vegetable found in southern China and parts of Southeast Asia. Studies have shown improper substrate hindered E. scandens seedling growth, causing water accumulation and nutrient deficiency. In pursuit of an ideal growth medium for E. scandens seedlings during the early stages, this study conducted a pot experiment to identify a mixed substrate with optimal water permeability and fertility. In this study, pure Alfisols soil treatment as the control (CK), and two soilless substrates (peat soil and perlite) were combined with Alfisols soil into different volume ratios, in order to better use soil resources from understory space and balance the texture of mixed substrates. The growth, physiological characteristics and nutrient status of 24-month-old E. scandens seedlings were determined after planting in different mixed ratios. The results showed that as the proportion of peat soil increased in the mix, most indexes exhibited an initial increase followed by a decline, while soluble protein content decreased consistently. Conversely, an increasing perlite ratio resulted in a general decline in most growth and physiological indexes. Root growth, biomass accumulation and chlorophyll content, peaked in the 66.67% Alfisols soil + 33.33% perlite (T4) treatment. Notably, T3 (66.67% Alfisols soil + 33.33% peat soil) showcased the best above-ground growth, while T1 (50.00% Alfisols soil + 50.00% peat soil) excelled in element content accumulation. In conclusion, the cultivation substrate should primarily consist of Alfisols soil, constituting at least 50%. The addition of peat soil enhances above-ground growth and nutrients accumulation, while perlite contributes to robust root development. One third of peat soil and a small amount of perlite can be added to the substrate during E. scandens seedling cultivation, and proper fertilization should also be used in order to increase nutrient accumulation in aboveground and underground parts. This research provides valuable insights into maximizing the potential of E. scandens seedlings through precise cultivation methods.

Substrate stiffness-dependent activation of Hippo pathway in cancer associated fibroblasts

The tumor microenvironment (TME) comprises a heterogenous cell population within a complex three-dimensional (3D) extracellular matrix (ECM). Stromal cells within this TME are altered by signaling cues from cancer cells to support uncontrolled tumor growth and invasion events. Moreover, the ECM also plays a fundamental role in tumor development through pathological remodeling, stiffening and interaction with TME cells. In healthy tissues, Hippo signaling pathway actively contributes to tissue growth, cell proliferation and apoptosis. However, in cancer, the Hippo signaling pathway is highly dysregulated, leading to nuclear translocation of the YAP/TAZ complex, which directly contributes to uncontrolled cell proliferation and tissue growth, and ECM remodeling and stiffening processes. Here, we compare the effect of increasing cell culture substrate stiffness, derived from tumor progression, upon the dysregulation of the Hippo signaling pathway in colorectal cancer-associated fibroblasts (CAFs) and normal colorectal fibroblasts (NFs). We correlate the dysregulation of Hippo pathway with the magnitude of the traction forces exerted by healthy and malignant stromal cells. We found that ECM stiffening is crucial in Hippo pathway dysregulation in CAFs, but not in normal fibroblasts.

Tracing potentially toxic elements in button mushroom cultivation and environmental implications: Insights via stable lead (Pb) isotope analysis

Elevated levels of potentially toxic elements (PTEs) have been frequently detected in substrates used for cultivating edible mushrooms. However, the distribution and accumulation pathways of PTEs within mushrooms and their cultivation substrates remain inadequately understood. This study specifically aims to clarify these aspects of button mushrooms (Agaricus bisporus). A systematic analysis was conducted on the distribution of PTEs (As, Cd, Cr, Cu, Ni, Pb, and Zn) and stable Pb isotope fingerprints across various components of the button mushroom cultivation systems, including the cultivation substrates (a mixture of wheat straw, chicken manure, soil, and additives), fruiting bodies (MR), and spent mushroom substrate (SMS). The methodology involved inductively coupled plasma mass‒spectrometry for PTE quantification and Pb isotope ratio analysis for source identification. The results demonstrated that wheat straw and additives were the main contributors to the endogenous accumulation of PTEs in MR, accounting for 35.0% and 33.2% of Pb accumulation, respectively. Furthermore, MR, wheat straw, and soil together contributed to more than 67% of the Pb loads in SMS. The study also revealed that the occurrence of PTEs in these substrates is predominantly driven by exogenous sources, particularly the significant contribution from anthropogenic activities. Overall, these findings provide valuable biogeochemical insights into the accumulation pathways and patterns of PTEs in button mushroom cultivation systems, with potential applications for cleaner food management and environmentally sustainable practices.

Essential Quality Attributes of Culture Media Used as Substrates in the Sustainable Production of Pre-Basic Potato Seeds

The sustainability of the primary sector is closely linked to meeting the demand for seeds using agro-industrial waste and bioresidues. Sustainability is a multidimensional concept focused on achieving environmental health, social justice, and economic viability. To this end, an experiment was designed based on a combination of biotechnological strategies accessible to many individuals. The first strategy involves the use of compost and vermicompost as cultivation substrates; the second is the in vitro acclimatization of potato plants to these substrates; and the third is the incorporation of Trichoderma asperellum into these substrates to determine the synergistic effect of both. The compost used in this work came from sewage sludge from an agri-food company (Cp); a dining room and pruning waste from a university campus (Cu); and vermicomposted coffee pulp waste (Cv). Each sample was mixed with coconut fiber (Fc) in proportions of 100, 75, 50, and 25%. In the resulting mixtures, María Bonita variety vitroplants were planted and placed in a greenhouse. The biometric response in the three cases indicated a dependence on the type of compost and the proportion of the coconut fiber mixture. The inoculation of Trichoderma asperellum with sewage sludge compost increased stem thickness (42.58%) and mini-tuber weight (6.74%). In contrast, uninoculated treatments showed the best performance in terms of the number of mini-tubers. A 50:50 mixture of sewage sludge compost with coconut fiber and without inoculation of Trichoderma asperellum was the best treatment for the production of pre-basic seeds of the María Bonita potato variety. The use of composted agricultural waste and bioresidues is shown as a valid and low-cost alternative for the sector, even independently of the incorporation of additional inoculants.

The influence of polyacrylamide gel substrate elasticity on primary cultures of rat retinal ganglion cells

In vitro primary cell culture models of retinal ganglion cells (RGC) are widely used to study pathomechanisms of diseases such as glaucoma. The biomechanic interaction with the culture substrate is known to influence core cellular functions. RGC cultures, however, are usually grown on rigid plastic or glass substrates. We hypothesized that soft polyacrylamide gel substrates may alter survival and neurite outgrowth of primary cultured RGC.Primary retinal cultures from postnatal (day 1-6) Wistar rats were grown on glass coverslips or polyacrylamide (PA) gel substrate with different Young’s elastic moduli (0.75, 10 or 30 kPa). Substrates were coated with Poly-l-lysine and / or laminin. RGC were immunostained with anti-beta-III-tubulin. Total neurite length, growth cone morphology, RGC density, mitochondrial morphology and transport as well as pro-survival pathways (Erk1/2, Akt, CREB) were assessed.PA gel substrates of E = 10 kPa significantly increased the total neurite length by factor 1.5 compared to glass (p = 0.02). The growth cone area was significantly larger by factor 5.3 on 30 kPa gels (p = 0.01). The presence of a substrate coating was more important for neurite outgrowth and RGC survival on PA gels (poly-l-lysine > laminin) than on glass. Neither mitochondrial morphology and motility nor the activation of pro-survival pathways significantly differed between the four substrates.PA gel substrates significantly enhanced RGC neurite outgrowth. The signaling cascades mediating this effect remain to be determined.

Developing a soft micropatterned substrate to enhance maturation of human induced pluripotent stem cell-derived cardiomyocytes

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSCCMs) offer numerous advantages as a biological model, yet their inherent immaturity compared to adult cardiomyocytes poses significant limitations. This study addresses hiPSCCM immaturity by introducing a physiologically relevant micropatterned substrate for long-term culture and maturation. An innovative microfabrication methodology combining laser etching and casting creates a micropatterned polydimethylsiloxane (PDMS) substrate with varying stiffness, from 2 to 50 kPa, mimicking healthy and fibrotic cardiac tissue. Platinum electrodes were integrated into the cell culture chamber enable pacing of cells at various frequencies. Subsequently, cells were transferred to the incubator for time-course analysis, ensuring contamination-free conditions. Cell contractility, cytosolic Ca2+ transient, sarcomere orientation, and nucleus aspect ratio were analyzed in a 2D hiPSCCM monolayer up to 90 days post-replating in relation to substrate micropattern dimensions. Culturing hiPSCCMs for three weeks on a micropatterned PDMS substrate (2.5–5 µm deep, 20 µm center-to-center spacing of grooves, 2–5 kPa stiffness) emerges as optimal for cardiomyocyte alignment, contractility, and cytosolic Ca2+ transient. The study provides insights into substrate stiffness effects on hiPSCCM contractility and Ca2+ transient at immature and mature states. Maximum contractility and fastest Ca2+transient kinetics occur in mature hiPSCCMs cultured for two to four weeks, with the optimum at three weeks, on a soft micropatterned PDMS substrate. MS proteomic analysis further revealed that hiPSCCMs cultured on soft micropatterned substrates exhibit advanced maturation, marked by significant upregulation of key structural, electrophysiological, and metabolic proteins. This new substrate offers a promising platform for disease modeling and therapeutic interventions. Statement of SignificanceHuman induced pluripotent stem cell derived cardiomyocytes (hiPSCCMs) have been transformative to disease-in-a-dish modeling, drug discovery and testing, and autologous regeneration for human hearts and their role will continue to expand dramatically. However, one of the major limitations of hiPSCCMs is that without intervention, the cells are immature and represent those in the fetal heart. We developed protocols for the fabrication of the PDMS matrices that includes variations in its stiffness and micropatterning. Growing our hiPSCCMs on matrices of comparable stiffness to a healthy heart (5 kPa) and grooves of 20 μm, generate heart cells typical of the healthy adult human heart.

Application and Development of Hydrogel in Soilless Culture

AbstractSoilless culture refers to the method of cultivating plants without soil as a growing medium, and the use of hydrogel in soilless culture overcomes many technical difficulties, but there are many applications that just stay at the experimental level, with fewer applications in fields. Although many new functionalized hydrogels may help to overcome the shortcomings of traditional media and give them more functionality, little research on hydrogels over the past decades has been reflected in soilless culture techniques. This paper details the concept and development of soilless culture, as well as several applications of hydrogels in soilless culture, including the use of hydrogels as soilless culture substrates and the application of hydrogel membranes. The important roles of different types of hydrogels in soilless culture, such as antimicrobial properties, salt tolerance and high water absorption, are highlighted, and these excellent properties largely overcome a series of drawbacks of traditional cultivation methods. The application of hydrogels in pest control is also discussed. Hydrogel applications to soilless culture have better prospects for the creation of more promising multifunctional composite hydrogel membranes and hydrogel substrates capable of supporting plants throughout their growth cycle.

Enhancing oyster mushroom cultivation by chickpea and wheat straw substrate for sustainable agriculture

This study aimed to explore the potential utilization of agro-lignocellulosic waste materials for oyster mushroom cultivation, addressing the challenge of managing waste associated with these materials. Various combinations of Chickpea Straw (CS) and Wheat Straw (WS) at different ratios-100% CS, 75% CS + 25% WS, 50% CS + 50% WS, 25% CS + 75% WS, and 100% WS-were investigated as substrates for mushroom cultivation. The experiment followed a ccompletely rrandomised design with four replications, monitoring developmental phases, yield, and biological efficiency (BE). Results indicated that using 100% WS as a substrate resulted in the fastest mycelium growth (spawn run), completing in an average of 14.50 days, 17.20 days from spawning to pinhead formation, and 20.60 days to first harvest, with the highest number of fruiting bodies produced. Chickpea straw contributed to the highest stipe width (1.52 cm), while WS 100% had the highest average yield (997.28 g) and BE (99.73%). A mixture of 75% CS and 25% WS showed promising results in terms of fruiting bodies, yield, and BE after 100% WS. Consequently, the study recommends the use of these substrates to optimize Pleurotus ostreatus cultivation yield.

Canola meal extract as a low-cost substrate in Mortierella alpina culture for the production of arachidonic acid

The demand for high-quality and cost-effective raw materials is growing across various industries. Canola meal (CM), a low-cost by-product of canola processing facilities, boasts an impressive nutritional profile comprising protein, carbohydrates, vitamins, and minerals, making it a potential candidate for microbial culture media. Saskatchewan leads globally in canola production and plans to expand its canola processing facilities. Commercial valorization of CM could not only optimize the management of this by-product but also offer additional incentives and financial returns to regional canola stakeholders. Efficient utilization of canola meal extract (CME) in producing polyunsaturated fatty acids (PUFAs), such as arachidonic acid (ARA), holds promise as valuable ingredients or base chemicals for the food, feed, and nutraceutical industries. This study focused on incorporating pH 10.5 treated alkaline extract (AE) and temperature 160°C treated subcritical extract (SE) of CM into the culture medium of Mortierella alpina, an oleaginous fungus, by replacing conventional yeast extract (YE). Trials investigated the effect of aeration levels ranging from 0.5 to 2.0 vvm and revealed that increasing oxygen supply significantly boosted fungal growth rate and ARA content in biomass up to 0.39 day⁻¹ and 21.86 %, respectively. ARA production correlated positively with biomass accumulation and lipid content. Replacing YE with CME resulted in less significant differences (P < 0.05) in biomass and lipid accumulation. Additionally, AE- and SE-supplemented media exhibited higher ARA accumulation of 144.91 mg/L and 165.8 mg/L, respectively, compared to standard media (124.87 mg/L). Therefore, this study introduces canola meal extract (CME) as a novel nitrogen source in microbial media, with comparative demonstrations showing its efficiency to be similar to that of conventional extracts.

Is Tea Waste A Promising Co-substrate for Optimizing The Cultivation, Growth, and Yield of Charleston Pepper (Capsicum annuum L.)?

To address growing concerns about sustainable agriculture and waste management, this study aimed to explore the viability of tea waste as an eco-friendly alternative substrate for cultivating Charleston peppers (Capsicum annuum), with the goal of optimizing plant growth and yield while reducing soil dependence, lowering cultivation costs, and repurposing agro-industrial waste. Six different substrate combinations were evaluated: 1) Tea waste, 2) Tea waste + Manure, 3) Tea waste + Soil, 4) Manure + Soil, 5) Tea waste + Manure + Soil, and 6) Tea waste + Manure + Soil + Perlite. Data were analyzed using both multivariate and univariate analyses to assess significant differences among treatments. Notably, significant differences in stem diameter were observed among plants grown on different substrates (one-way MANOVA, p &amp;lt;.05). However, plant height and chlorophyll content remained unaffected by substrate type. Although leaf structure exhibited considerable variation across treatments, no significant difference in dry matter content was observed. These results demonstrate that tea waste, especially when combined with other materials, is a promising sustainable substrate for Charleston pepper cultivation, potentially reducing soil dependence and agro-industrial waste.